WO2023004941A1 - 用于治疗失眠的丝素蛋白微针透皮贴片及其制备方法 - Google Patents

用于治疗失眠的丝素蛋白微针透皮贴片及其制备方法 Download PDF

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WO2023004941A1
WO2023004941A1 PCT/CN2021/117078 CN2021117078W WO2023004941A1 WO 2023004941 A1 WO2023004941 A1 WO 2023004941A1 CN 2021117078 W CN2021117078 W CN 2021117078W WO 2023004941 A1 WO2023004941 A1 WO 2023004941A1
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microneedle
silk fibroin
solution
melatonin
additive
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PCT/CN2021/117078
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English (en)
French (fr)
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祁珍珍
卢神州
曹嘉欣
陶晓晟
胡昂辉
郭仕杰
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苏州大学
帕拉森医药生物技术(上海)有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A61K31/4045—Indole-alkylamines; Amides thereof, e.g. serotonin, melatonin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A61K47/40—Cyclodextrins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/20—Hypnotics; Sedatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0046—Solid microneedles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0015—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0053—Methods for producing microneedles
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention belongs to the technical field of transdermal administration, and in particular relates to a silk fibroin microneedle transdermal patch for treating insomnia and a preparation method thereof.
  • Melatonin originally discovered in the bovine pineal gland in 1958, is a hormone synthesized by the pineal gland and released at night that has been shown to phase change or retime circadian rhythms and sleep-wake schedules.
  • Administration of derived melatonin The physiological role of melatonin in regulating seasonal and circadian rhythms is thought to be mediated by a specific, high-affinity, G protein-coupled cell membrane receptor.
  • Oral melatonin enters the liver through the digestive tract, leading to the first-pass effect of the liver, resulting in a decrease in the bioavailability of the drug, which can only reach about 20%.
  • oral melatonin is difficult to reach a certain blood concentration quickly and maintain a high concentration for a long time. Therefore, transdermal delivery of melatonin may be a good route of administration.
  • microneedle transdermal drug delivery system is a new type of transdermal drug delivery system, which is a needle-shaped complex structure with a size of micron.
  • microneedles Compared with other transdermal drug delivery methods, microneedles have unique advantages: 1Compared with chemical osmosis and iontophoresis, microneedles can pierce the stratum corneum to create tiny pores for the entry of macromolecular drugs; 2microneedles The channel through which the needle pierces the human body is only on the micron scale, without damage to the skin, and not enough to touch the nerve endings, so there is almost no pain; 3The combination of microneedles and other methods can further improve the transdermal penetration of drugs.
  • the Chinese invention patent with the application number CN201811017105.2 discloses a method for preparing a melatonin microneedle patch, which includes the following steps: pouring an aqueous solution of a polymer material containing melatonin onto a microneedle patch with a microneedle hole matrix mold, and the polymer material aqueous solution is filled in the micropores on the mold, after the mold is pasted with a prefabricated backing layer, the backing layer is attached to the melatonin micropores formed in the micropores The back side of the needle is then frozen, thawed, peeled off and dried to obtain the melatonin microneedle patch.
  • its release rate is slow, and the elimination rate of the drug in the body is fast.
  • the present invention aims to provide a silk fibroin microneedle transdermal patch for treating insomnia and its preparation method.
  • Melatonin can be delivered through the skin, which can quickly reach a certain level of blood drug, and maintain a relatively high level of drug in the body for a long time. High concentration.
  • the preparation method of the silk fibroin microneedle transdermal patch for treating insomnia comprises the following steps,
  • step S3 adding the aqueous solution of the microneedle matrix modification additive obtained in step S1 and the melatonin solution obtained in step S2 to the silk fibroin aqueous solution with a concentration of 20-60 mg/mL to obtain a mixed solution I;
  • step S4 Add the aqueous drug release additive solution obtained in step S1 to the mixed solution I to obtain the mixed solution II, the concentration of melatonin in the mixed solution II is 0.5-20 mg/mL, and the concentration of the microneedle matrix modification additive is 4 -25mg/mL, the concentration of the drug release additive is 0.05-10mg/mL;
  • microneedle base modification additive is one or more of proline, sorbitol and xylitol.
  • the concentration of the aqueous solution of the microneedle matrix modification additive is 100-1000 mg/mL, preferably 300-900 mg/mL.
  • the mass ratio of the microneedle matrix modification additive to the silk fibroin is 1-5:10.
  • the drug dissolution additive is one or more of isopropanol, n-propanol, n-butanol, isobutanol and 2-butanol.
  • the concentration of the aqueous solution of the drug dissolution additive is 100-800 mg/mL, preferably 300-800 mg/mL.
  • the mass ratio of drug dissolution additive to silk fibroin is 1:5-20.
  • the drug release additive is one or more of ⁇ -cyclodextrin, hydroxypropyl- ⁇ -cyclodextrin and ⁇ -cyclodextrin.
  • the concentration of the aqueous solution of the drug release additive is 10-100 mg/mL.
  • the concentration of melatonin in the melatonin solution is 5-60 mg/mL, preferably 15-55 mg/mL.
  • both the melatonin solution and the mixed solution II are stored at 2-8° C. and protected from light.
  • the silk fibroin aqueous solution is obtained from silkworm silk by degumming, dissolving, and dialysis purification.
  • the concentration of the silk fibroin aqueous solution is 30-50 mg/mL.
  • the concentration of melatonin in the mixed solution II is 5-15 mg/mL
  • the concentration of the microneedle matrix modification additive is 10-25 mg/mL
  • the concentration of the drug release additive is 2-10 mg/mL.
  • the concave hole of the microneedle mold is conical, with a length of 200-800 ⁇ m and a bottom diameter of 100-400 ⁇ m.
  • microneedle mold is made of silicone rubber.
  • step S5 the drying is carried out under the condition of constant temperature and humidity (20-30°C, 55-65%) and air circulation for 4-8h. Since the drug-dissolving additive is a small molecule alcohol, the drug will Dissolving additives are all volatilized and removed.
  • Another aspect of the present invention provides the silk fibroin microneedle transdermal patch for treating insomnia prepared by the above preparation method.
  • the microneedle body of the obtained silk fibroin microneedle transdermal patch is solid conical, the length of the needle body is 200-800 ⁇ m, the diameter of the bottom is 100-400 ⁇ m, and the interval between adjacent needle bodies is 0.2-2 mm.
  • silk fibroin As a natural protein, silk fibroin has good mechanical properties and plasticity, good compatibility with human tissue, biodegradable, non-toxic, and easy to process and prepare.
  • the large number of hydrophilic groups it contains can better contain water-soluble drugs, break through the lipid-rich stratum corneum, and improve the ability of drugs to permeate through the skin.
  • Melatonin is a fat-soluble drug, insoluble in water, and difficult to mix evenly with silk fibroin solution.
  • the intervention of drug-dissolving additives makes melatonin change the molecular three-dimensional arrangement of the unit cell in the original crystal structure, so that the molecular state can enter the drug-dissolving additives. It can then be evenly blended with silk fibroin to form uniform microneedles. Melatonin is evenly distributed inside the microneedles, and a uniform release rate is obtained in the subsequent transdermal drug release.
  • the addition of the microneedle matrix modification additive is to promote the formation of a certain crystal structure of the silk fibroin, reduce the rate of dissolution and loss, form a certain swelling pore size, and control the release speed of melatonin.
  • the addition of drug release additives is to accelerate the release rate of melatonin and increase the cumulative release of melatonin. Due to the high hydrophilicity of the drug release additive, it is more conducive to the rapid dissolution and release of the drug in the body, and a higher blood drug concentration can be achieved in a short time. At the same time, the solubility of melatonin in body fluids is improved, the bioavailability of melatonin is further improved, and the drug effect is enhanced, thereby reducing the amount of drug added. Therefore, after the drug release additive is included with the drug, the ratio, type, and form of the drug can affect the solubility of the drug in the inclusion compound.
  • the inclusion of melatonin by cyclodextrin can play four kinds of controlled release effects, namely immediate release, delayed release (time-controlled release), extended release and controlled release. Selecting the appropriate cyclodextrin, adding the appropriate amount, and rationally designing the formulation of melatonin microneedles can improve or regulate the release rate.
  • the technical solution of the present invention has the following advantages: the melatonin microneedle patch prepared by the present invention can quickly reach a certain blood drug level, and maintain a relatively high concentration of the drug in the body for a long time. At the same time, the microneedle transdermal patch avoids the liver first-pass effect and can improve the bioavailability of melatonin.
  • FIG. 1 is a microscope photo of the microneedle patch prepared in Example 1 of the present invention.
  • Fig. 2 is a graph showing the plasma-drug concentration curve of the microneedle patch prepared in Example 1 of the present invention used in rats.
  • Fig. 3 is an in vitro transdermal drug release curve of the microneedle patch prepared in Examples 1-6 of the present invention.
  • Preparation of silk fibroin solution Weigh 3g NaHCO 3 and 1g Na 2 CO 3 and dissolve in 4000mL deionized water, heat to boiling, put 80g silk, keep at 98°C for 30min, take it out and wash it in deionized water. After repeating the above steps three times, place in a 60°C oven to dry to obtain silk fibers. Prepare 9.3mol/L LiBr aqueous solution, take 100mL LiBr aqueous solution and heat it to 65°C in a water bath, put 15g of silk fiber in several times and stir to dissolve, then continue to heat and stir for 40min.
  • the liquid was put into a dialysis bag and placed in deionized water for dialysis for 72 hours, and the deionized water was replaced every 2 hours. After dialysis, the solution is filtered to obtain a silk fibroin solution. Take a small amount of solution to dry and weigh to calculate the solution mass fraction.
  • Preparation of small molecule additive aqueous solution weigh proline and add it to deionized water, shake and mix evenly, so that the concentration of proline in the aqueous solution is 500 mg/mL; weigh hydroxypropyl- ⁇ -cyclodextrin and add it to deionized water, Shake and mix to make the concentration of hydroxypropyl- ⁇ -cyclodextrin in the aqueous solution 100 mg/mL; measure isopropanol, add it to deionized water, shake and mix to make the concentration of isopropanol in the aqueous solution 600 mg/mL.
  • Preparation of melatonin solution Weigh melatonin into a centrifuge tube, add 1 mL of isopropanol aqueous solution, shake and mix, so that the concentration of melatonin in the solution is 50 mg/mL.
  • Preparation of the mixed solution slowly add the above proline aqueous solution and melatonin solution into the silk fibroin aqueous solution while stirring, the concentration of the silk fibroin aqueous solution is 50 mg/mL; then add the aqueous hydroxypropyl- ⁇ -cyclodextrin solution, Obtain the mixed solution; Make the concentration of melatonin in the mixed solution be 15mg/mL, proline concentration be 10mg/mL, hydroxypropyl-beta-cyclodextrin be 5mg/mL, place under 4 °C after thorough mixing. light save.
  • Injection molding of the solution Take 3mL of the mixed solution and pour it into the silicone rubber microneedle mold, and vacuum 3 times to remove the bubbles in the solution and the microneedle template, so that the solution can fully enter the cavity of the needle body.
  • Drying of the microneedle patch place the defoamed mold system in a constant temperature and humidity room (25°C, 65%) under air circulation for 8 hours, and demould to obtain a silk fibroin microneedle transdermal patch containing melatonin piece.
  • the microscopic photo of the obtained silk fibroin microneedle transdermal patch is shown in Figure 1. It can be seen that the length of the microneedle is 600 ⁇ m, and the diameter of the bottom surface is about 300 ⁇ m.
  • the plasma-drug concentration curve of the obtained silk fibroin microneedle transdermal patch applied to rats is shown in FIG. 2 . It can be seen from the figure that the time to reach the maximum blood drug concentration in rats is 0.41 ⁇ 0.03h, and the maximum blood drug concentration is 10.08 ⁇ 0.02ng/mL. In the time range of 6 hours, the drug concentration in rats can maintain the effective drug concentration level and remain stable. It shows that the microneedle patch prepared by the present invention can reach a certain blood drug concentration in a relatively short period of time, covering the entire sleep cycle, and achieving the effect of assisting sleep.
  • Preparation of small molecule additive aqueous solution Weigh sorbitol and add it to deionized water, shake and mix until the concentration of sorbitol in the aqueous solution is 400 mg/mL; weigh hydroxypropyl- ⁇ -cyclodextrin and add it to deionized water, shake and mix Evenly, so that the concentration of hydroxypropyl- ⁇ -cyclodextrin in the aqueous solution is 80mg/mL. Measure n-propanol, add it to deionized water, shake and mix, so that the concentration of n-propanol in the aqueous solution is 500 mg/mL.
  • Preparation of melatonin solution Weigh melatonin into a centrifuge tube, first add 0.8mL n-propanol aqueous solution, shake and mix well, so that the concentration of melatonin in the solution is 40mg/mL, and keep the melatonin solution at 4°C. light save.
  • Preparation of the mixed solution slowly add the above-mentioned sorbitol aqueous solution and melatonin solution into the silk fibroin aqueous solution while stirring, the concentration of the silk fibroin aqueous solution is 40 mg/mL; then add the hydroxypropyl- ⁇ -cyclodextrin aqueous solution to obtain Mix the solution; make the concentration of melatonin in the mixed solution 8 mg/mL, sorbitol 10 mg/mL, and hydroxypropyl- ⁇ -cyclodextrin 5 mg/mL, mix well and store in the dark at 4°C .
  • Injection molding of the solution Take 3mL of the mixed solution and pour it into the silicone rubber microneedle mold, and vacuum 3 times to remove the bubbles in the solution and the microneedle template, so that the solution can fully enter the cavity of the needle body.
  • Drying of the microneedle patch place the defoamed mold system in a constant temperature and humidity room (25°C, 65%) under air circulation for 8 hours, and demould to obtain a silk fibroin microneedle transdermal patch containing melatonin piece.
  • Preparation of small molecule additive aqueous solution Weigh sorbitol and add it to deionized water, oscillate and mix to make the concentration of sorbitol in the aqueous solution 600 mg/mL; weigh ⁇ -cyclodextrin and add it to deionized water, oscillate and mix to make the aqueous solution The concentration of ⁇ -cyclodextrin in the medium is 50mg/mL. Measure n-butanol, add it to deionized water, shake and mix, so that the concentration of n-butanol in the aqueous solution is 600mg/mL.
  • Preparation of melatonin solution Weigh melatonin into a centrifuge tube, first add 1mL n-butanol aqueous solution, shake and mix well, so that the concentration of melatonin in the solution is 60mg/mL, and keep the melatonin solution at 4°C in the dark save.
  • Preparation of the mixed solution slowly add the above-mentioned sorbitol aqueous solution and melatonin solution into the silk fibroin aqueous solution while stirring, the concentration of the silk fibroin aqueous solution is 50 mg/mL; then add the hydroxypropyl- ⁇ -cyclodextrin aqueous solution to obtain Mix the solution; make the concentration of melatonin in the mixed solution 7 mg/mL, the concentration of sorbitol 20 mg/mL, and the concentration of ⁇ -cyclodextrin 5 mg/mL, mix well and store at 4°C in the dark.
  • Injection molding of the solution Take 3mL of the mixed solution and pour it into the silicone rubber microneedle mold, and vacuum 3 times to remove the bubbles in the solution and the microneedle template, so that the solution can fully enter the cavity of the needle body.
  • Drying of the microneedle patch place the defoamed mold system in a constant temperature and humidity room (25°C, 65%) under air circulation for 8 hours, and demould to obtain a silk fibroin microneedle transdermal patch containing melatonin piece.
  • Preparation of small molecule additive aqueous solution weigh proline and add it to deionized water, oscillate and mix evenly, so that the concentration of proline in the aqueous solution is 700 mg/mL; weigh ⁇ -cyclodextrin and add it to deionized water, oscillate and mix evenly, The concentration of ⁇ -cyclodextrin in the aqueous solution was 30 mg/mL. Measure n-butanol, add it to deionized water, shake and mix, so that the concentration of n-butanol in the aqueous solution is 600mg/mL.
  • Preparation of melatonin solution Weigh melatonin into a centrifuge tube, first add 1.5mL n-butanol aqueous solution, shake and mix well, so that the concentration of melatonin in the solution is 30mg/mL, and keep the melatonin solution at 4°C. light save.
  • Preparation of the mixed solution slowly add the above proline aqueous solution and melatonin solution into the silk fibroin aqueous solution while stirring, the concentration of the silk fibroin aqueous solution is 40 mg/mL; then add the aqueous hydroxypropyl- ⁇ -cyclodextrin solution, Obtain a mixed solution; make the concentration of melatonin in the mixed solution be 12 mg/mL, the concentration of proline be 15 mg/mL, and the concentration of ⁇ -cyclodextrin be 3 mg/mL, mix well and store in the dark at 4°C .
  • Injection molding of the solution Take 3mL of the mixed solution and pour it into the silicone rubber microneedle mold, and vacuum 3 times to remove the bubbles in the solution and the microneedle template, so that the solution can fully enter the cavity of the needle body.
  • Drying of the microneedle patch place the defoamed mold system in a constant temperature and humidity room (25°C, 65%) under air circulation for 8 hours, and demould to obtain a silk fibroin microneedle transdermal patch containing melatonin piece.
  • Preparation of small molecule additive aqueous solution Weigh proline into deionized water, oscillate and mix evenly, so that the concentration of proline in the aqueous solution is 900 mg/mL; weigh ⁇ -cyclodextrin into deionized water, oscillate and mix, The concentration of ⁇ -cyclodextrin in the aqueous solution was 70 mg/mL. Measure isobutanol, add it to deionized water, shake and mix, so that the concentration of isobutanol in the aqueous solution is 800mg/mL.
  • Preparation of melatonin solution Weigh melatonin into a centrifuge tube, first add 2mL isobutanol aqueous solution, shake and mix well, so that the concentration of melatonin in the solution is 55mg/mL, and keep the melatonin solution at 4°C in the dark save.
  • Preparation of the mixed solution slowly add the above proline aqueous solution and melatonin solution into the silk fibroin aqueous solution while stirring, the concentration of the silk fibroin aqueous solution is 50 mg/mL; then add the aqueous hydroxypropyl- ⁇ -cyclodextrin solution, Obtain a mixed solution; make the concentration of melatonin in the mixed solution be 15 mg/mL, the concentration of proline be 25 mg/mL, and the concentration of ⁇ -cyclodextrin be 10 mg/mL, mix well and store in the dark at 4°C .
  • Injection molding of the solution Take 3mL of the mixed solution and pour it into the silicone rubber microneedle mold, and vacuum 3 times to remove the bubbles in the solution and the microneedle template, so that the solution can fully enter the cavity of the needle body.
  • Drying of the microneedle patch place the defoamed mold system in a constant temperature and humidity room (25°C, 65%) under air circulation for 8 hours, and demould to obtain a silk fibroin microneedle transdermal patch containing melatonin piece.
  • Preparation of small molecule additive aqueous solution weigh xylitol and add it to deionized water, oscillate and mix evenly, so that the concentration of xylitol in the aqueous solution is 300mg/mL; weigh ⁇ -cyclodextrin and add it to deionized water, oscillate and mix evenly, The concentration of ⁇ -cyclodextrin in the aqueous solution was 10 mg/mL. Measure 2-butanol, add it to deionized water, shake and mix, so that the concentration of 2-butanol in the aqueous solution is 300 mg/mL.
  • Preparation of melatonin solution Weigh melatonin in a centrifuge tube, first add 1.5mL of 2-butanol aqueous solution, shake and mix, so that the concentration of melatonin in the solution is 15mg/mL, and the melatonin solution 4 °C and protected from light.
  • Preparation of the mixed solution slowly add the above xylitol aqueous solution and melatonin solution into the silk fibroin aqueous solution while stirring, the concentration of the silk fibroin aqueous solution is 50 mg/mL; then add the hydroxypropyl- ⁇ -cyclodextrin aqueous solution, A mixed solution was obtained; the concentration of melatonin in the mixed solution was 5 mg/mL, the concentration of xylitol was 15 mg/mL, and the concentration of ⁇ -cyclodextrin was 2 mg/mL. After being fully mixed, it was stored at 4° C. in the dark.
  • Injection molding of the solution Take 3mL of the mixed solution and pour it into the silicone rubber microneedle mold, and vacuum 3 times to remove the bubbles in the solution and the microneedle template, so that the solution can fully enter the cavity of the needle body.
  • Drying of the microneedle patch place the defoamed mold system in a constant temperature and humidity room (25°C, 65%) under air circulation for 8 hours, and demould to obtain a silk fibroin microneedle transdermal patch containing melatonin piece.
  • the in vitro transdermal drug release curves of the microneedle patches prepared in Examples 1-6 are shown in Figure 3. It can be seen that by adding different types of microneedle matrix modification additives, different swelling degrees can be obtained. Silk fibroin microneedles, thereby adjusting the speed of drug release in the microneedles and the time for drug release to reach equilibrium. Drug dissolution additives can enhance the dissolution of drugs, so as to obtain microneedles with different drug loadings. The drug release additive can promote the release of the drug in the microneedle, improve the cumulative drug release rate of the microneedle and the bioavailability of melatonin.

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Abstract

一种用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，包括以下步骤：分别配制微针基体改性添加剂、药物溶解添加剂和药物释放添加剂的水溶液；向药物溶解添加剂水溶液中加入褪黑素，得到褪黑素溶液；将微针基体改性添加剂水溶液和褪黑素溶液加入丝素蛋白水溶液中，再加入药物释放添加剂水溶液，得到混合溶液II；将混合溶液II倒入微针模具中，经抽真空脱泡、干燥、脱模，制得所述用于治疗失眠的丝素蛋白微针透皮贴片。所述制备方法得到的褪黑素微针贴片能够快速达到一定的血药水平，并且长时间保持体内药物的较高浓度；同时，微针透皮贴片避免了肝脏首过效应，能够提高褪黑素的生物利用率。

Description

用于治疗失眠的丝素蛋白微针透皮贴片及其制备方法 技术领域
本发明属于经皮给药技术领域，具体涉及一种用于治疗失眠的丝素蛋白微针透皮贴片及其制备方法。
背景技术
褪黑素最初于1958年在牛松果体中发现，是一种由松果体合成并在夜间释放的激素，已显示具有相变或重定时生物钟节律和睡眠-苏醒时间表作用的是外源性褪黑素给药。褪黑素在调节季节性和昼夜节律的生理作用被认为是一种特异性、高亲和力的和G蛋白偶联的细胞膜受体介导的。目前，研究最多的是口服途径。口服褪黑素经过消化道进入肝脏，导致肝脏首过效应，造成药物的生物利用度下降，只能达到20％左右。同时，口服褪黑素难以较快达到一定的血药浓度和维持长时间的高浓度。因此，褪黑素的透皮递送可能是给药的好途径。
微针透皮给药系统是透皮给药系统一种新型方式，它是尺寸为微米级，呈针状的复杂结构。与其他透皮给药方法相比，微针有其独到的优势：①与化学促渗，离子导入相比，微针能够刺破角质层产生微小的孔道以供大分子药物的进入；②微针刺破人体的通道仅微米级，对皮肤无损伤，且不足以触及神经末梢，所以几乎无痛觉；③微针与其他方法联用，可进一步提高药物透皮渗透能力。
现有技术中，申请号为CN201811017105.2的中国发明专利公开了一种褪黑素微针贴剂的制备方法，其包括如下步骤：将含有褪黑素的聚合物材料水溶液浇注于带有微孔矩阵的模具上，并使所述聚合物材料水溶液填充于模具上的微孔内，在模具贴附预制的背衬层后，使背衬层帖附于微孔内形成的褪黑素微针的背面，然后进行冷冻、解冻后，揭膜、干燥，得到所述褪黑素微针贴剂。但是其释放速度慢，药物在体内的消除速度快。
发明内容
本发明旨在提供一种用于治疗失眠的丝素蛋白微针透皮贴片及其制备方法，褪黑素透皮递送，能够快速达到一定的血药水平，并且长时间保持体内药物的较高浓度。
按照本发明的技术方案，所述用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，包括以下步骤，
S1：分别配制微针基体改性添加剂水溶液、药物溶解添加剂水溶液和药物释放添加剂水溶液；
S2：向所述药物溶解添加剂水溶液中加入褪黑素，混匀得到褪黑素溶液；
S3：将步骤S1所得微针基体改性添加剂水溶液和步骤S2所得褪黑素溶液加入浓度为20-60mg/mL的丝素蛋白水溶液中，得到混合溶液I；
S4：向所述混合溶液I中加入步骤S1所得药物释放添加剂水溶液，得到混合溶液II，所述混合溶液II中褪黑素的浓度为0.5-20mg/mL，微针基体改性添加剂浓度为4-25mg/mL，药物释放添加剂的浓度为0.05-10mg/mL；
S5：将所述混合溶液II倒入微针模具中，经抽真空脱泡、干燥、脱模，制得所述用于治疗失眠的丝素蛋白微针透皮贴片。
进一步的，所述微针基体改性添加剂为脯氨酸、山梨醇和木糖醇中的一种或多种。
进一步的，所述微针基体改性添加剂水溶液的浓度为100-1000mg/mL，优选浓度为300-900mg/mL。
进一步的，所述混合溶液II中，微针基体改性添加剂与丝素蛋白的质量比为1-5：10。
进一步的，所述药物溶解添加剂为异丙醇、正丙醇、正丁醇、异丁醇和2-丁醇中的一种或多种。
进一步的，所述药物溶解添加剂水溶液的浓度为100-800mg/mL，优选浓度为300-800mg/mL。
进一步的，所述混合溶液II中，药物溶解添加剂与丝素蛋白的质量比为1：5-20。
进一步的，所述药物释放添加剂为α-环糊精、羟丙基-β-环糊精和γ-环糊精中的一种或多种。
进一步的，所述药物释放添加剂水溶液的浓度为10-100mg/mL。
进一步的，所述褪黑素溶液中褪黑素的浓度为5-60mg/mL，优选浓度为15-55mg/mL。
进一步的，褪黑素溶液和混合溶液II均置于2-8℃下避光保存。
进一步的，所述丝素蛋白水溶液由家蚕蚕丝通过脱胶，溶解，透析纯化获得。
进一步的，所述丝素蛋白水溶液的浓度为30-50mg/mL。
优选的，所述混合溶液II中褪黑素的浓度为5-15mg/mL，微针基体改性添加剂浓度为10-25mg/mL，药物释放添加剂的浓度为2-10mg/mL。
进一步的，所述微针模具的凹孔为圆锥形，长度为200-800μm，底部直径为100-400μm。
进一步的，微针模具采用硅橡胶材质。
进一步的，步骤S5中，干燥是在恒温恒湿间(20-30℃、55-65％)空气流通的条件下干燥4-8h，由于药物溶解添加剂为小分子醇类，在干燥过程中药物溶解添加剂全部挥发除去。
本发明的另一方面提供了上述制备方法制得的用于治疗失眠的丝素蛋白微针透皮贴片。所得丝素蛋白微针透皮贴片的微针针体为实心圆锥状，针体的长度为200-800μm，底部直径为100-400μm，相邻针体之间的间隔为0.2-2mm。
发明原理：
丝素蛋白作为一种天然蛋白质，具有良好的机械性能及可塑性，与人体组织相容性好，可生物降解，无毒害，易于加工制备。利用它含有的大量亲水基团可更好地与水溶性药物相包合，突破富含脂类的角质层，提高药物经皮渗透 的能力。
褪黑素为脂溶性药物，难溶于水，难以与丝素蛋白溶液混合均匀。药物溶解添加剂的介入，使褪黑素改变了原来晶体结构中晶胞的分子立体排列，得以分子状态进入到药物溶解添加剂中。然后能够与丝素蛋白均匀共混，形成均匀的微针。褪黑素均匀分布在微针的内部，在其后的透皮释药中获得均匀的释放速度。微针基体改性添加剂的加入，是为了促使丝素蛋白形成一定的结晶结构，降低溶解溶失率，形成一定的溶胀孔径，控制褪黑素的释药速度。
药物释放添加剂的加入，是为了加快褪黑素的释放速度，提高褪黑素的累积释放量。由于药物释放添加剂的高亲水性，更有利于药物在体内快速溶解释放，短时间内能达到较高的血药浓度。同时提高褪黑素在体液中的溶解度，进一步提高褪黑素的生物利用度，增强药效，从而减少药物添加量。故药物释放添加剂与药物包合后，前者的比例、种类、以及药物的存在形式都可以影响包合物中药物的溶解度。同时，对于褪黑素，通过环糊精对褪黑素的包合作用可以起到4种控制释放作用，分别为即时释放、延迟释放(时间控制释放)、延长释放和控制释放。选择合适的环糊精，适当的添加量，合理设计褪黑素微针的配方，可以改善或调控释放速度。
本发明的技术方案相比现有技术具有以下优点：本发明制备的褪黑素微针贴片能够快速达到一定的血药水平，并且长时间保持体内药物的较高浓度。同时，微针透皮贴片避免了肝脏首过效应，能够提高褪黑素的生物利用率。
附图说明
图1为本发明实施例1所制备的微针贴片的显微镜照片。
图2为本发明实施例1所制备的微针贴片用于大鼠的血浆-药物浓度曲线图。
图3是本发明实施例1-6所制备的微针贴片的体外透皮释药曲线图。
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明，以使本领域的技术人员可以更好地理解本发明并能予以实施，但所举实施例不作为对本发明的限定。
制备例1
丝素蛋白溶液的制备：称取3g NaHCO 3和1g Na 2CO 3溶于4000mL去离子水中，加热至沸腾，放入80g蚕丝，保持98℃微沸30min，取出后于去离子水清洗。重复上述步骤三次后放置于60℃烘箱中干燥得到丝素纤维。配制9.3mol/L的LiBr水溶液，取100mL的LiBr水溶液水浴加热至65℃，分多次放入丝素纤维15g搅拌溶解后，继续加热并搅拌40min。液体装入透析袋中放置于去离子水中透析72h，每隔2h更换去离子水。透析后溶液经过过滤获得丝素蛋白溶液。取少量溶液烘干称量计算溶液质量分数。
实施例1
小分子添加剂水溶液的制备：称取脯氨酸加入去离子水中，震荡混匀，使水溶液中脯氨酸的浓度为500mg/mL；称取羟丙基-β-环糊精加入去离子水中，震荡混匀，使水溶液中羟丙基-β-环糊精浓度为100mg/mL；量取异丙醇，加入去离子水中，震荡混匀，使水溶液中异丙醇的浓度为600mg/mL。
褪黑素溶液的制备：称取褪黑素于离心管中，加入1mL异丙醇水溶液，震荡混匀，使溶液中褪黑素的浓度为50mg/mL。
混合溶液的制备：将上述脯氨酸水溶液与褪黑素溶液一边搅拌一边缓慢加入丝素蛋白水溶液中，丝素蛋白水溶液浓度为50mg/mL；再加入羟丙基-β-环糊精水溶液，得到混合溶液；使得混合溶液中褪黑素的浓度为15mg/mL，脯氨酸浓度为10mg/mL，羟丙基-β-环糊精为5mg/mL，充分混合后置于4℃下避光保存。
溶液的注模：取3mL混合溶液倒在硅橡胶微针模具中，抽真空3次，去除溶液和微针模板中的气泡，使得溶液充分进入针体空腔。
微针贴片的干燥：将脱泡后的模具系统置于恒温恒湿间(25℃、65％)空气流通处下干燥8h，脱模得到含褪黑素的丝素蛋白微针透皮贴片。
所得丝素蛋白微针透皮贴片的显微镜照片如图1所示，可以看到微针长度在600μm，底面直径在300μm左右。
所得丝素蛋白微针透皮贴片用于大鼠的血浆-药物浓度曲线图如图2所示。由图可知，大鼠体内达到最大血药浓度的时间为0.41±0.03h，最大血药浓度为10.08±0.02ng/mL。在6h的时间范围内，大鼠体内的药物浓度能够维持有效药物浓度水平并保持稳定。说明本发明制备的微针贴片能够在较短时间内达到一定的血药浓度，涵盖整个睡眠周期，达到辅助随眠的功效。
实施例2
小分子添加剂水溶液的制备：称取山梨醇加入去离子水中，震荡混匀，使水溶液中山梨醇的浓度为400mg/mL；称取羟丙基-β-环糊精加入去离子水中，震荡混匀，使水溶液中羟丙基-β-环糊精浓度为80mg/mL。量取正丙醇，加入去离子水中，震荡混匀，使水溶液中正丙醇的浓度为500mg/mL。
褪黑素溶液的制备：称取褪黑素于离心管中，先加入0.8mL正丙醇水溶液，震荡混匀，使溶液中褪黑素的浓度为40mg/mL，褪黑素溶液4℃避光保存。
混合溶液的制备：将上述山梨醇水溶液与褪黑素溶液一边搅拌一边缓慢加入丝素蛋白水溶液中，丝素蛋白水溶液浓度为40mg/mL；再加入羟丙基-β-环糊精水溶液，得到混合溶液；使得混合溶液中褪黑素的浓度为8mg/mL，山梨醇浓度为10mg/mL，羟丙基-β-环糊精为5mg/mL，充分混合后置于4℃下避光保存。
溶液的注模：取3mL混合溶液倒在硅橡胶微针模具中，抽真空3次，去除溶液和微针模板中的气泡，使得溶液充分进入针体空腔。
微针贴片的干燥：将脱泡后的模具系统置于恒温恒湿间(25℃、65％)空气流通处下干燥8h，脱模得到含褪黑素的丝素蛋白微针透皮贴片。
实施例3
小分子添加剂水溶液的制备：称取山梨醇加入去离子水中，震荡混匀，使水溶液中山梨醇的浓度为600mg/mL；称取α-环糊精加入去离子水中，震荡混匀，使水溶液中α-环糊精浓度为50mg/mL。量取正丁醇，加入去离子水中，震荡混匀，使水溶液中正丁醇的浓度为600mg/mL。
褪黑素溶液的制备：称取褪黑素于离心管中，先加入1mL正丁醇水溶液，震荡混匀，使溶液中褪黑素的浓度为60mg/mL，褪黑素溶液4℃避光保存。
混合溶液的制备：将上述山梨醇水溶液与褪黑素溶液一边搅拌一边缓慢加入丝素蛋白水溶液中，丝素蛋白水溶液浓度为50mg/mL；再加入羟丙基-β-环糊精水溶液，得到混合溶液；使得混合溶液中褪黑素的浓度为7mg/mL，山梨醇浓度为20mg/mL，α-环糊精的浓度为5mg/mL，充分混合后置于4℃下避光保存。
溶液的注模：取3mL混合溶液倒在硅橡胶微针模具中，抽真空3次，去除溶液和微针模板中的气泡，使得溶液充分进入针体空腔。
微针贴片的干燥：将脱泡后的模具系统置于恒温恒湿间(25℃、65％)空气流通处下干燥8h，脱模得到含褪黑素的丝素蛋白微针透皮贴片。
实施例4
小分子添加剂水溶液的制备：称取脯氨酸加入去离子水中，震荡混匀，使水溶液中脯氨酸的浓度为700mg/mL；称取α-环糊精加入去离子水中，震荡混匀，使水溶液中α-环糊精浓度为30mg/mL。量取正丁醇，加入去离子水中，震荡混匀，使水溶液中正丁醇的浓度为600mg/mL。
褪黑素溶液的制备：称取褪黑素于离心管中，先加入1.5mL正丁醇水溶液，震荡混匀，使溶液中褪黑素的浓度为30mg/mL，褪黑素溶液4℃避光保存。
混合溶液的制备：将上述脯氨酸水溶液与褪黑素溶液一边搅拌一边缓慢加入丝素蛋白水溶液中，丝素蛋白水溶液浓度为40mg/mL；再加入羟丙基-β-环糊精水溶液，得到混合溶液；使得混合溶液中褪黑素的浓度为12mg/mL，脯氨酸浓度为15mg/mL，α-环糊精的浓度为3mg/mL，充分混合后置于4℃下避光保存。
溶液的注模：取3mL混合溶液倒在硅橡胶微针模具中，抽真空3次，去除溶液和微针模板中的气泡，使得溶液充分进入针体空腔。
微针贴片的干燥：将脱泡后的模具系统置于恒温恒湿间(25℃、65％)空 气流通处下干燥8h，脱模得到含褪黑素的丝素蛋白微针透皮贴片。
实施例5
小分子添加剂水溶液的制备：称取脯氨酸加入去离子水中，震荡混匀，使水溶液中脯氨酸的浓度为900mg/mL；称取γ-环糊精加入去离子水中，震荡混匀，使水溶液中γ-环糊精浓度为70mg/mL。量取异丁醇，加入去离子水中，震荡混匀，使水溶液中异丁醇的浓度为800mg/mL。
褪黑素溶液的制备：称取褪黑素于离心管中，先加入2mL异丁醇水溶液，震荡混匀，使溶液中褪黑素的浓度为55mg/mL，褪黑素溶液4℃避光保存。
混合溶液的制备：将上述脯氨酸水溶液与褪黑素溶液一边搅拌一边缓慢加入丝素蛋白水溶液中，丝素蛋白水溶液浓度为50mg/mL；再加入羟丙基-β-环糊精水溶液，得到混合溶液；使得混合溶液中褪黑素的浓度为15mg/mL，脯氨酸浓度为25mg/mL，γ-环糊精的浓度为10mg/mL，充分混合后置于4℃下避光保存。
溶液的注模：取3mL混合溶液倒在硅橡胶微针模具中，抽真空3次，去除溶液和微针模板中的气泡，使得溶液充分进入针体空腔。
微针贴片的干燥：将脱泡后的模具系统置于恒温恒湿间(25℃、65％)空气流通处下干燥8h，脱模得到含褪黑素的丝素蛋白微针透皮贴片。
实施例6
小分子添加剂水溶液的制备：称取木糖醇加入去离子水中，震荡混匀，使水溶液中木糖醇的浓度为300mg/mL；称取γ-环糊精加入去离子水中，震荡混匀，使水溶液中γ-环糊精浓度为10mg/mL。量取2-丁醇，加入去离子水中，震荡混匀，使水溶液中2-丁醇的浓度为300mg/mL。
褪黑素溶液的制备：称取褪黑素于离心管中，先加入1.5mL的2-丁醇水溶液，震荡混匀，使溶液中褪黑素的浓度为15mg/mL，褪黑素溶液4℃避光保存。
混合溶液的制备：将上述木糖醇水溶液与褪黑素溶液一边搅拌一边缓慢加入丝素蛋白水溶液中，丝素蛋白水溶液浓度为50mg/mL；再加入羟丙基-β-环糊 精水溶液，得到混合溶液；使得混合溶液中褪黑素的浓度为5mg/mL，木糖醇浓度为15mg/mL，γ-环糊精为2mg/mL，充分混合后置于4℃下避光保存。
溶液的注模：取3mL混合溶液倒在硅橡胶微针模具中，抽真空3次，去除溶液和微针模板中的气泡，使得溶液充分进入针体空腔。
微针贴片的干燥：将脱泡后的模具系统置于恒温恒湿间(25℃、65％)空气流通处下干燥8h，脱模得到含褪黑素的丝素蛋白微针透皮贴片。
检测实施例1
实施例1-6所制备的微针贴片的体外透皮释药曲线图如图3所示，可以看出，通过添加不同种类比例的微针基体改性添加剂，可以获得具有不同溶胀度的丝素蛋白微针，从而调节微针中药物释放的速度和药物释放达到平衡的时间。药物溶解添加剂能够增强药物的溶解量，从而获得不同载药量的微针。药物释放添加剂可以促进微针中药物的释放量，提高微针的累计释药率以及褪黑素的生物利用度。
显然，上述实施例仅仅是为清楚地说明所作的举例，并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引申出的显而易见的变化或变动仍处于本发明创造的保护范围之中。

Claims (12)

  1. 一种用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，包括以下步骤，
    S1：分别配制微针基体改性添加剂水溶液、药物溶解添加剂水溶液和药物释放添加剂水溶液；
    S2：向所述药物溶解添加剂水溶液中加入褪黑素，混匀得到褪黑素溶液；
    S3：将步骤S1所得微针基体改性添加剂水溶液和步骤S2所得褪黑素溶液加入浓度为20-60mg/mL的丝素蛋白水溶液中，得到混合溶液I；
    S4：向所述混合溶液I中加入步骤S1所得药物释放添加剂水溶液，得到混合溶液II，所述混合溶液II中褪黑素的浓度为0.5-20mg/mL，微针基体改性添加剂浓度为4-25mg/mL，药物释放添加剂的浓度为0.05-10mg/mL；
    S5：将所述混合溶液II倒入微针模具中，经抽真空脱泡、干燥、脱模，制得所述用于治疗失眠的丝素蛋白微针透皮贴片。
  2. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述微针基体改性添加剂为脯氨酸、山梨醇和木糖醇中的一种或多种。
  3. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述微针基体改性添加剂水溶液的浓度为100-1000mg/mL。
  4. 如权利要求1、2或4所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述混合溶液II中，微针基体改性添加剂与丝素蛋白的质量比为1-5：10。
  5. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述药物溶解添加剂为异丙醇、正丙醇、正丁醇、异丁醇和2-丁醇中的一种或多种。
  6. 如权利要求1或5所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备 方法，其特征在于，所述药物溶解添加剂水溶液的浓度为100-800mg/mL。
  7. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述药物释放添加剂为α-环糊精、羟丙基-β-环糊精和γ-环糊精中的一种或多种。
  8. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述药物释放添加剂水溶液的浓度为10-100mg/mL。
  9. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述混合溶液II中，药物释放添加剂与丝素蛋白的质量比为1：5-20。
  10. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述褪黑素溶液中褪黑素的浓度为5-60mg/mL。
  11. 如权利要求1所述的用于治疗失眠的丝素蛋白微针透皮贴片的制备方法，其特征在于，所述微针模具的凹孔为圆锥形，长度为200-800μm，底部直径为100-400μm。
  12. 一种如权利要求1-11中任一项所述的制备方法制得的用于治疗失眠的丝素蛋白微针透皮贴片。
PCT/CN2021/117078 2021-07-30 2021-09-08 用于治疗失眠的丝素蛋白微针透皮贴片及其制备方法 WO2023004941A1 (zh)

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